Abstract

Studies have shown that embryonic stem (ES) cells can be successfully differentiated into liver cells, which offer the potential unlimited cell source for a variety of end-stage liver disease. In our study, in order to induce mouse ES cells to differentiate into hepatocyte-like cells under chemically defined conditions, ES cells were induced by sodium butyrate as inducer for 7 days in the first phase, followed by the combination of hepatocyte growth factor and dexamethasone as inducers for 12 days in the second phase based on an adherent culture system with a two-step induction. The results from the morphology, gene expression, protein molecular markers and cell function of ES-D3 cells derived hepatocyte-like cells demonstrated that mES cells can be differentiated efficiently in vitro to functional hepatocytes under chemically defined conditions, which might be useful as an in vitro system for hepatocyte transplantation therapy and toxicity screening in drug discovery.

Key words: Embryonic stem cells, hepatic-like cells, in vitro differentiation, sodium butyrate, hepatocyte growth factor, dexamethason.

Abbreviation

mES, Mouse embryonic stem cell; AFP, α-fetoprotein; AAT, α-1-antitrypsin; HNF, hepatocyte nuclear factor; TAT, tyrosine aminotransferase; LIF,leukemia inhibitory factor; EBs, embryoid bodies; BAL, bioartificial liver; CK,cytokeratin; DAPI, 4,6-diamidino-2-phenylindole; Dex, dexamethasone; EGF,epidermal growth factor; FITC, fluorescein isothiocyanate; HGF, hepatocyte growth factor; ICG, indocyanine green; iPSC, induced pluripotent stem cell; MEF, mouse embryonic fibroblast; NEAA, non-essential amino acid; OLT, orthotopic liver transplantation; PAS, periodic acid Schiff.